Hair cells in the frog semicircular canal, studied by the whole-cell patch-clamp technique, display three distinct Ca2+ currents: two non-inactivating components (L type and R type, the latter termed R2 in the following) and a second R type current (termed R1), which runs down first and inactivates in a Ca2+-dependent fashion. Since intracellular EGTA, up to 5 mM, did not display major effects on such inactivation, we used increasing amounts of BAPTA in the patch pipette, to control [Ca2+]i more efficiently and investigate whether modifications in [Ca2+]i at the cytoplasmic side of the channel affect the inactivation of the RI component and in general the gating of all channel types. The results here reported show that (1) K+ currents heavily contaminate recordings obtained using high concentrations of BAPTA in its commercially available K+ salt form; (2) BAPTA Cs+ salt can be satisfactorily employed to obtain reliable recordings; (3) the kinetics of channel gating and R1-channel inactivation are indeed markedly affected by effectively buffering [Ca2+]i.

Ca2+-dependent kinetics of hair cell Ca2+ currents resolved with the use of cesium BAPTA

RISPOLI, Giorgio;MARTINI, Marta;ROSSI, Marialisa;RUBBINI, Gemma;
2000

Abstract

Hair cells in the frog semicircular canal, studied by the whole-cell patch-clamp technique, display three distinct Ca2+ currents: two non-inactivating components (L type and R type, the latter termed R2 in the following) and a second R type current (termed R1), which runs down first and inactivates in a Ca2+-dependent fashion. Since intracellular EGTA, up to 5 mM, did not display major effects on such inactivation, we used increasing amounts of BAPTA in the patch pipette, to control [Ca2+]i more efficiently and investigate whether modifications in [Ca2+]i at the cytoplasmic side of the channel affect the inactivation of the RI component and in general the gating of all channel types. The results here reported show that (1) K+ currents heavily contaminate recordings obtained using high concentrations of BAPTA in its commercially available K+ salt form; (2) BAPTA Cs+ salt can be satisfactorily employed to obtain reliable recordings; (3) the kinetics of channel gating and R1-channel inactivation are indeed markedly affected by effectively buffering [Ca2+]i.
Rispoli, Giorgio; Martini, Marta; Rossi, Marialisa; Rubbini, Gemma; Fesce, R.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/516679
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